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1 Tabel privind lucrările rezultate în urma tezei de doctorat, publicate sau în curs de publicare în reviste BDI/ISI Titlul Autori Determination of content in almond milk by colorimetrically assay with Folin Ciocalteu Reagent Dan Alina Florentina, Anca Maria Tifrea, TitaOvidiu Nr. autori Nume revista BuletinAgir ISBN ISSN -798 Anul publicarii Tip revista (BDI/ISI) Denumire BDI/Factor de impact ISI Link la zona publica a publicatiei Publicatia (in format PDF) BDI INDEX COPERNICUS INTERNATIONAL
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3 DETERMINATION OF PROTEIN CONTENT IN ALMOND MILK BY COLORIMETRICALLY ASSAY WITH FOLIN CIOCALTEAU REAGENT Eng.drd. Alina Florentina DAN Faculty of Agricultural Sciences, Food Industry and Environment Protection, Lucian Blaga University of Sibiu Eng.drd.Anca Maria ȚIFREA Faculty of Agricultural Sciences, Food Industry and Environment Protection, Lucian Blaga University of Sibiu Ph.D.eng. Ovidiu TITA Department of Agricultural Sciences and Food Engineering, Faculty of Agricultural Sciences, Food Industry and Environment Protection, Lucian Blaga University of Sibiu REZUMAT. Scopul studiului a fost determinarea conţinutului total de e din laptele de migdale prin utilizarea unei metode mai putin sensibile la medicamente care interferează şi sunt mai sensibile la e. Două tipuri de probe de lapte de migdale au fost amestecate cu Reactiv /Lowry. Amestecul a fost incubat la temperatura camerei timp de minute. După ce reactivul Folin-Ciocalteau a fost adăugat şi amestecul a fost incubat timp de alte de minute, conţinutul total de ă a fost analizat colorimetric. Absorbanţa a fost măsurată pe un spectrofotometru (Shimadzu) la 7 nm. Curba de etalonare s-a realizat folosind o substanţă de referinţă dupa cum urmeaza:, mg / ml;, mg / ml;, mg / ml;, mg / ml;, mg / ml. Rezultatele pentru conţinutul total de e din lapte de migdale s-au situat în jurul valorii de,7%. Cuvinte cheie: lapte de migdale, e, Folin-Ciocalteau. ABSTRACT. The aim of this study was to determinate the total content of in almond milk by using a procedure less sensitive to interfering agents and more sensitive to. In order to do this, two types of samples of almond milk were mixed with Reagent /Lowry reagent. The mixture was incubated at room temperature for minutes. After the Folin-Ciocalteau reagent was added and the mixture was incubated for another minutes, the total content of was assayed colorimetrically. The absorbance was measured on a spectrophotometer (Shimadzu Scientific Instruments) at 7 nm. The calibration curve was made using a reference substance as followed:, mg/ml;, mg/ml;, mg/ml;, mg/ml;, mg/ml. The results for total content of from almond milk were around,7 %. Keywords: almond milk,, Folin-Ciocalteau. INTRODUCTION Almond milk is a milky drink made from ground alomnds. One of his most important benefits comes from the fact that it does not contain lactose. Some of the people are intolerant to this sugar and if they consume animal milk thay can present some of the following symptoms: bloating, diarrhea, discomfort, vomiting. Researchers showed that consuming almond milk instead of animal milk, the behavior of people have visibly improved and those symptoms disappeared( Iaconoa G, 8). The high content of monounsaturated fatty acids from almond milk has been asociated with reducing heart disease and the big amount of vitamin E with antioxidant activity(jenkins,). It is low in calories and carbohydrates, being advantageous for those trying to lose weight. It is known that content from almond is high. The purpose of this study was to determinate the content of s from an almond milk obtained from Spanish almonds milk using a method that doesn t requires long time and, in the same time, being less sensitive to interfering agents.. MATERIALS AND METHODS.Chemicals and reagents. The following analytical reagents were obtained from Sigma Co: sodium carbonate, cupric sulfate, sodium potassium tartrate, sodium lauryl sulfate (SLS),sodium hydroxide and Folin Ciocalteau 9
4 SIPA, November -. Nyiregyháza, Hungary reagent. materials for were obtained also from sigma Co. The copper reagent was prepared as followed: mg sodium carbonate were disolved in ml water,, mg cupric sulfate in ml water and, mg sodium potassium tartrate in ml water. The three solution were mixed. For preparing % SLS solution, mg was disolved in ml water and for preparing M solution of NaOH, mg were disolved in ml water. Lowry reagent was prepared by mixing sodium lauryl sulfate, % with sodium hydroxide, M and cooper reagent in these proportions: ::(Stoescheck,99) The Folin Ciocalteau reagent,, N was prepared by mixing ml N Folin with 9 ml Water and kept in an amber bottle.. Colorimetricaly assay of total content of g almond milk, respectively g of almond milk with olive oil were weight in a ml wolumetric flask and brought to volume with water. Because the concentration could not be estimated, a range of - dilutions were prepared. The intermediate dilution (: ). was taken in calculations. Briefly, ml almond milk(indicated dilution), respectively ml of almond milk with olive oil, and ml Lowry reagent were mixed and incubated at room temparature for minutes. Then,, ml Folin Ciocalteau were added, vortexed and the mixture were incubated for minutes more at room temperature (Lowry,9). The absorbance was measured on a spectrophotometer (Shimadzu Scientific Instruments) at 7 nm. For reading absorbance, glass cuvetts were used.the calibration curve was made using a reference substance as followed:, mg/ml;, mg/ml;, mg/ml;, mg/ml;, mg/ml.. RESULTS AND DISCUSSION Fourteen samples of two types of almond milk have been investigated: almond milk normally and almond milk with olive oil. The values for reference substance concentration and its absorbance are shown in table and figure nuber shows the calibration curve, where it can be seen that the value of R-squared is,998. Table. Substance concentration and its absorbance Concentration Absorbance substance, of reference Mean, SD %RSD Readings [nm] [mg/ml],8,,,87,89,8 absorbance, nm,8,,,,,89,7,8,,8,,,,99,,,,9,,,,7,8,,,, concentration, mg /ml,89,9,87,,,8,99,9,98,89,9,9,7,7,7 y =,8x +,8 R =,998 Fig. Calibration curve and standard equation for reference substance Table.. Almond milk samples and their total content of Total content of Total content of Sample determinated from determinated number almond milk with olive from almond oil,% milk,% Sample,8, Sample,8, Sample,8,7 Sample,77, Sample,79, Sample,8, Sample 7,78, Sample 8,8, Sample 9,78,7 Sample,8,9 Sample,8, Sample,8,7 Sample,8, Sample,8, Table shows the values determined with calorimetrically assay with Folin Ciocalteau reagent for every sample. The values are expressed as percentage. One of the statistical method used for data analysis was Summary (Microsoft Excel-Analyze-it, figure and figure ). This analysis shows the number of observations analysed and summary statistics.
5 The 7 th International Conference INTEGRATED SYSTEMS FOR AGRI-FOOD PRODUCTION F r e q u e n c y 7 Histogram,77,78,79,8,8,8,8 Groups - total content of from almond milk, % Normal Fit (Mean=.8, SD=.8) Fig.. Histogram for total content of from almond milk, % As it can be seen from figure number, the mean value for total content of from almond milk were,8%. The most values were between,8 and,8 %. Regarding total content of from almond milk with olive oil, figure number shows that the most values were between, and, %. The mean value for content in this case was,98 %. The results showed a insignificant difference between the two type of almond milk. It can be seen that the total content of from almond milk with olive oil was smaller than the value of total content of from normaly almond milk. This can be due to interfering s with almond milk fats during the procedure. F r e q u e n c y Histogram,,,,,,, Groups - total content of from almond milk with olive oil,% Normal Fit (Mean=.98, SD=.7) Fig.. Histogram for total content of from almond milk with olive oil, % In order to compare the values of the two groups t-test(median test) was used as a statistical method (Microsoft Excel-Analyze-it, Table ). The test showed that the mean difference is,79, insignificant for this types of almond milk. Tabel. T-test applied to compare the values of the two groups. n 8 Groups n Mean SE SD Total,8,8,8 content of from almond milk, % Total,98,9,7 content of from almond milk with olive oil, % Mean,79 difference 9% CI, to,9 SE, t statistic, DF, -tailed p <, Recording absorbance for this method needed only be done within minutes of each for this procedure. This procedure demonstrated to be less sensitive to interfering agents and more sensitive to.. ACKNOWLEDGMENT This study was made possible through the project "Romanian Research Integration in European Research Context, POSDRU/88/./S/7", financed from European Social Fund by Operational Sectoral Programme for Human Resources Development 7-.. REFERENCES [] Iaconoa, G., Lospallutia, M.L., Licastroa, G., Scalicia, C., A new formula based on almond milknext term for management of cow previous termmilknext term intolerance and food allergies, Gastroenterologia Pediatrica, Palermo, 8 [] Jenkins,D.J., Kendall, C.W., Marchie, A., Parker, T.L., Connelly, P.W., Qian, W., Haight, J.S., Faulkner, D., Vidgen, E., Lapsley, K.G., Spiller, G.A., Dose response of almonds on coronary heart disease risk factors: blood lipids, oxidized low-density lipos, lipo(a), homocysteine, and pulmonary nitric oxide: a randomized, controlled, crossover trial., Circulation, Journal of the American Heart Association, ;():7-, [] Lowry,OH,NJ Rosbrough,AL Farr, and RJ Randall. J.Biol.Chem. 9:, 9 [] Stoescheck CM.Quantitation of Protein. Methods in Enzimology 8: -9, 99 Determination of total content of from almond milk by colorimetrically assay with Folin Ciocalteau reagent proved to be a quick method.
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